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Acid Phosphatase (ACP) Activity Assay Kit (PNPP Method)
SKU: E-BC-K010-M-500
Acid Phosphatase (ACP) Activity Assay Kit (PNPP Method)
| SKU # | E-BC-K010-M |
| Detection Instrument | Microplate reader (400-415 nm, optimum wavelength: 405 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | ALP |
| Sample Type | Serum, plasma, tissue |
| Sensitivity | 0.2 U/L |
| Detection Range | 0.2–50 U/L |
| Detection Method | Colorimetric method |
| Assay Time | 30 min |
| Precision | Inter-assay CV: 4.5% | intra-assay CV: 2.8% |
| Other Reagents Required | PBS (0.01 M, pH 7.4) |
| Storage | This product can be stored at -20°C for 12 months with shading light. |
| Valid period | 12 months |
Dilution Of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
|---|---|
| 10% Epipremnum aureum tissue homogenate | 5-10 |
| Mouse plasma | 5-10 |
| Rat plasma | 5-10 |
| Human urine | 1 |
| Human plasma | 5-10 |
| 10% Rat spleen tissue homogenate | 20-30 |
| 10% Rat liver tissue homogenate | 20-30 |
| 10% Rat kidney tissue homogenate | 20-30 |
The diluent is PBS (0.01 M, pH 7.4). For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Disodium p-nitrobenzene phosphate (p-NPP), a widely used phosphatase chromogenic substrate, can form p- nitrophenol under the action of acid phosphatase. Under alkaline conditions, p-nitrophenol is yellow and has a maximum absorption peak at 405 nm. The darker of the yellow product is, the higher of the ACP activity is. Therefore, the activity of ACP can be calculated by measuring the OD value at 405 nm.
Kit Components & Storage
| Item | Component | Size 1(96 T) | Size 2 (500 Assays) | Storage |
| Reagent 1 | Buffer Solution | 20 mL ×1 vial | 50 mL ×2 vials | -20℃, 12 months |
| Reagent 2 | Substrate | Powder×3 vials | Powder ×15 vials | -20℃, 12 months, shading light |
| Reagent 3 | Standard | Powder × 1 vial | Powder × 5 vials | 20℃, 12 months shading light |
| Reagent 4 | Chromogenic Agent | 24 mL × 1 vial | 60 mL × 2 vials | -20℃, 12 month |
| Microplate | 96 wells | No requirement | ||
| Plate Sealer | 2 pieces |
Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.
Technical Data:
Parameter:
Intra-assay Precision
Three human serum samples were assayed in replicates of 20 to determine precision within an assay (CV = Coefficient of Variation).
| Parameters | Sample 1 | Sample 2 | Sample 3 |
|
Mean (U/L) |
0.50 | 15.00 | 38.00 |
| %CV | 3.2 | 2.6 | 2.6 |
Inter-assay Precision
Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
|
Mean (U/L) |
0.50 | 15.00 | 38.00 |
| %CV | 4.3 | 4.7 | 4.5 |
Sensitivity
The analytical sensitivity of the assay is 0.2 U/L ACP. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.
Standard Curve:
As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:
| Concentration (mmol/L) | 0 | 0.05 | 0.1 | 0.2 | 0.25 | 0.3 | 0.4 | 0.5 |
| Average OD | 0.039 | 0.145 | 0.469 | 0.469 | 0.577 | 0.675 | 0.892 | 1.121 |
| Absoluted OD | 0 | 0.105 | 0.214 | 0.429 | 0.538 | 0.626 | 0.852 | 1.081 |